Chlamydomonas cilia proteins

Axonemal dyneins are highly complex molecular motors that power ciliary motility. These multi-subunit enzymes are assembled at dedicated sites within the cytoplasm. At least nineteen cytosolic factors are specifically needed for the generation of dynein holoenzymes and/or their trafficking to the growing cilium. Many proteins are subject to N-terminal processing and acetylation which can generate degrons subject to the AcN-end rule and alter N-terminal electrostatics, generate new binding interfaces, and affect subunit stoichiometry through targeted degradation. Here we have used mass spectrometry of cilia samples and electrophoretically purified dynein heavy chains from Chlamydomonas to define their N-terminal processing; we also detail the N-terminal acetylase complexes present in this organism. We identify four classes of dynein heavy chain based on their processing pathways by two distinct acetylases one of which is dependent on methionine aminopeptidase activity. In addition, we fi..., These data were generated from whole cilia and electrophoretically purified dynein heavy chains digested with trypsin or Asp-N using an Orbitrap Eclipse mass spectrometer and positive mode electrospray ionization. Data were searched against the Chlamydomonas v.6.1 proteome. , , # Chlamydomonas cilia proteins [https://doi.org/10.5061/dryad.mw6m90635](https://doi.org/10.5061/dryad.mw6m90635) These are scaffold files that contain the proteomic data for whole cilia and electrophoretically purified axonemal dynein heavy chains from Chlamydomonas. ## Description of the data and file structure Four scaffold files are uploaded. 1. \"Whole Cilia\" - Chlamydomonas cilia digested with trypsin or endoproteinase AspN. 2. \"Dynein Heavy Chains\" - Electrophoretically purified wildtype Chlamydomonas axonemal dynein heavy chains digested with trypsin. 3. \"Dynein Heavy Chains 2\" - Electrophoretically purified wildtype Chlamydomonas axonemal dynein heavy chains digested with trypsin. 4. \"AspN-digested Dynein Heavy Chains: - Electrophoretically purified wildtype and oda2 Chlamydomonas axonemal dynein heavy chains digested with endoproteinase AspN.轴丝动力蛋白（Axonemal dyneins）是一类高度复杂的分子马达，负责驱动纤毛运动。这类多亚基酶在细胞质内的特定位点完成组装。生成动力蛋白全酶（holoenzyme）并将其转运至正在生长的纤毛，至少需要19种特异性胞质因子。众多蛋白质会经历N端加工与乙酰化修饰，该修饰过程可产生遵循AcN端规则的降解信号（degrons），改变N端区域的静电特性，形成全新的结合界面，并通过靶向降解影响亚基的化学计量比。 本研究针对衣藻（Chlamydomonas）来源的纤毛样本开展质谱分析，并对电泳纯化的动力蛋白重链进行表征，明确了其N端加工过程；同时详细解析了该物种中存在的N端乙酰化酶复合物。我们基于两种不同乙酰化酶的加工途径，将动力蛋白重链分为四类，其中一类乙酰化酶的活性依赖于甲硫氨酸氨肽酶。此外，本研究的实验数据源自经胰蛋白酶（trypsin）或Asp-N内切酶消化的全纤毛样本与电泳纯化的动力蛋白重链，检测采用Orbitrap Eclipse质谱仪与正模式电喷雾电离。质谱数据的检索基于衣藻v6.1版本蛋白质组。 # 衣藻纤毛蛋白质组 [https://doi.org/10.5061/dryad.mw6m90635](https://doi.org/10.5061/dryad.mw6m90635) 本数据集以支架文件形式存储衣藻全纤毛样本与电泳纯化的轴丝动力蛋白重链的蛋白质组学数据。 ## 数据与文件结构说明 本次共上传4个支架文件： 1. "全纤毛样本"：经胰蛋白酶或Asp-N内切酶消化的衣藻纤毛样本。 2. "动力蛋白重链"：经电泳纯化的野生型衣藻轴丝动力蛋白重链，经胰蛋白酶消化。 3. "动力蛋白重链2"：经电泳纯化的野生型衣藻轴丝动力蛋白重链，经胰蛋白酶消化。 4. "Asp-N酶切动力蛋白重链"：经电泳纯化的野生型与oda2突变型衣藻轴丝动力蛋白重链，经Asp-N内切酶消化。